Much progress has been made in the electrokinetic phenomena inside nanochannels in the last decades. As the dimensions of the nanochannels are compatible to that of the electric double layer (EDL), the electrokinetics inside nanochannels indicate many unexpected behaviors, which show great potential in the fields of material science, biology, and chemistry. This review summarizes the recent development of nanofluidic electrokinetics in both fundamental and applied research. First, the techniques for constructing nanochannels are introduced to give a guideline for choosing the optimal fabrication technique based on the specific feature of the nanochannel. Then, the theories and experimental investigations of the EDL, electroosmotic flow, and electrophoresis of nanoparticles inside the nanochannels are discussed. Furthermore, the applications of nanofluidic electrokinetics in iontronics, sensing, and biomolecule separation fields are summarized. In Section 5, some critical challenges and the perspective on the future development of nanofluidic electrokinetics are briefly proposed.